Analysis of Protein Posttranslational Modifications Using DIGE-Based Proteomics

  • Robert M. DeKroon
  • Jennifer B. Robinette
  • Cristina Osorio
  • Joseph S. Y. Jeong
  • Eric Hamlett
  • Mihaela Mocanu
  • Oscar AlzateEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 854)


Difference gel electrophoresis (DIGE) is most often used to assess relative changes in the expression levels of individual proteins in multiple complex samples, and this information is valuable in making inferences about relative protein activity. However, a protein’s activity is not solely dependent upon its expression level. A change in activity may also be influenced by myriad posttranslational modifications (PTMs), including palmitoylation, ubiquitination, oxidation, and phosphorylation. In this chapter, we describe the use of DIGE to determine specific PTMs by introducing specific labels or changes in pI and/or molecular weight.

Key words

Difference gel electrophoresis Neuroproteomics Oxidation Palmitoylation Phosphory-lation Posttranslational modification Ubiquitination 



This work was supported by the UNC Systems-Proteomics Center and the Duke Neuroproteomics Center.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Robert M. DeKroon
    • 1
  • Jennifer B. Robinette
    • 2
  • Cristina Osorio
    • 2
  • Joseph S. Y. Jeong
    • 1
  • Eric Hamlett
    • 1
  • Mihaela Mocanu
    • 2
  • Oscar Alzate
    • 1
    Email author
  1. 1.Department of Cell and Developmental BiologyUniversity of North CarolinaChapel HillUSA
  2. 2.UNC Systems-Proteomics Center, Program of Molecular Biology and Biotechnology, School of MedicineUniversity of North CarolinaChapel HillUSA

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